Project Summary Alcohol consumption contributes to approximately 6% of worldwide deaths and is a major cause of morbidity and mortality within the United States. These statistics support a pressing need for understanding the biochemical mechanisms underlying alcohol toxicity and the pathogenesis of alcohol-associated liver disease (ALD). Chronic alcohol metabolism impacts numerous cellular pathways including glycolysis, lipid metabolism, the TCA cycle as well as antioxidant and inflammatory responses. There is a known biochemical link between metabolic alterations, oxidative stress, and protein thiol redox switches (e.g., cysteine (Cys) residues); however, very little information exists regarding how chronic alcohol metabolism impacts hepatic thiol redox signaling and control networks. Our preliminary data supports the notion that alcohol metabolism, protein acetylation, and Cys redox are highly associated. Therefore, we present an innovative approach for investigating how alcohol metabolism impacts thiol redox signaling and control. Central to our aims, the thiol redox proteome is an adaptive interface that provides a means to sense, avoid, and defend against oxidants and other toxicants. Therefore, the hypothesis of this proposal is that alcohol metabolism impacts thiol redox signaling and control through lysine acetylation, resulting in hepatic dyshomeostasis and contributing to ALD. We will investigate the proposed specific aims to test our hypothesis: Specific Aim 1: Characterize altered thiol redox signaling and control due to alcohol metabolism. Specific Aim 2: Utilize Sirtuin 1 overexpression to define mechanisms of acetylation-redox signaling and control. Specific Aim 3: Integrate mechanisms of redox- Cys and acetyl-Lys to elucidate CoAlation specific redox signaling. We will execute these research aims utilizing a cutting-edge proteomics and bioinformatics approach to reveal novel redox sensing mechanisms within hepatocytes. Elucidating how alcohol metabolism alters hepatic redox signaling and control through novel post-translational modifications will support the development of targeted clinical interventions to ameliorate ALD in millions of Americans.